Agilent's spectral flow cytometry system distinguishes similar fluorochromes
Agilent Technologies has introduced a new spectral flow cytometry system capable of differentiating fluorochromes with highly overlapping spectra, increasing flexibility in multi-color panel design.
Agilent Technologies, Inc. has demonstrated the capability of its NovoCyte Opteon spectral flow cytometer to distinguish between fluorochromes with highly similar spectral signatures. This technology addresses a key limitation in traditional flow cytometry.
Unlike conventional methods that rely on compensation to correct for fluorescence spillover, spectral flow cytometry utilizes spectral unmixing. This process employs mathematical algorithms to break down the spectral signature of a multicolor sample, allowing for the effective separation of fluorochromes based on their unique spectral characteristics, even those with near-identical peak emissions.
The company reported successful differentiation of FITC and BB515, fluorochromes with a similarity index of 0.99, on the NovoCyte Opteon. The system also demonstrated the ability to resolve other similarly spectrally overlapping combinations, including BV421/Pacific Blue and APC/Alexa Fluor 647.
This advancement expands the options for researchers in designing multi-color flow cytometry panels. Agilent notes that while spectral unmixing resolves similar fluorochromes, spillover spreading cannot be entirely eliminated and requires careful consideration in analyses.